Monarch DNA Gel Extraction Kit Protocol

Add 4 volumes of ethanol to one volume of DNA Wash Buffer according to kit size. For a 50-prep kit, add 20 ml ethanol to 5 ml buffer; for a 250-prep kit, add 100 ml ethanol to 25 ml buffer.

Visualize the DNA fragment under UV light and excise it from the gel using a razor blade or scalpel, cutting as close to the DNA as possible to minimize excess agarose and UV exposure.

Transfer the excised band to a 1.5 ml microcentrifuge tube, weigh it, and add 4 volumes of Monarch Gel Dissolving Buffer. Incubate at 37-55°C for 5-10 minutes, mixing periodically until the agarose completely dissolves.

Load the dissolved sample onto the column and close the cap. Centrifuge at 16,000 x g for 1 minute and discard the flow-through; DNA is now bound to the column.

Re-insert the column into a collection tube and add 200 μl of DNA Wash Buffer. Centrifuge for 1 minute and discard flow-through. Repeat the wash step with another 200 μl of DNA Wash Buffer and centrifuge again.

Transfer the column to a clean microfuge tube ensuring the column tip does not contact the flow-through. Add 6 or more μl of DNA Elution Buffer to the column membrane center and incubate for 1 minute to maximize yield.

Centrifuge the column for 1 minute at 16,000 x g and collect the flow-through in the tube below, which contains your final gel-extracted DNA.